GuiLu-ErXian Glue extract promotes mesenchymal stem cells (MSC)-Induced chondrogenesis via exosomes release and delays aging in the MSC senescence process

Yong-Hong Yang; Che-Sheng Wen; Yung-Ling Kuo; Su-Ling Fu; Tung-Yi Lin; Chao-Ming Chen; Po-Kuei Wu; Wei-Ming Chen; Jir-You Wang

doi:10.1016/j.jep.2023.116784

GuiLu-ErXian Glue extract promotes mesenchymal stem cells (MSC)-Induced chondrogenesis via exosomes release and delays aging in the MSC senescence process

J Ethnopharmacol. 2023 Dec 5:317:116784. doi: 10.1016/j.jep.2023.116784. Epub 2023 Jun 14.

Authors

Yong-Hong Yang¹, Che-Sheng Wen², Yung-Ling Kuo³, Su-Ling Fu⁴, Tung-Yi Lin⁵, Chao-Ming Chen⁶, Po-Kuei Wu⁷, Wei-Ming Chen⁸, Jir-You Wang⁹

Affiliations

¹ Institute of Traditional Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC. Electronic address: lucas258012@gmail.com.
² Department of Orthopedics, Cheng-Hsin General Hospital, Taipei, Taiwan, ROC. Electronic address: jasonfennywen@gmail.com.
³ School of Chinese Medicine for Post Baccalaureate, College of Medicine, I-Shou University, Kaohsiung, Taiwan, ROC. Electronic address: ylkylk@yahoo.com.tw.
⁴ Institute of Traditional Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC. Electronic address: slfu@nycu.edu.tw.
⁵ Institute of Traditional Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC. Electronic address: biotungyi@gmail.com.
⁶ Department of Orthopedics, Taipei Veterans General Hospital, Taipei, Taiwan, ROC; Department of Orthopedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, ROC; Institute of Clinical Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC. Electronic address: excelnova@gmail.com.
⁷ Department of Orthopedics, Taipei Veterans General Hospital, Taipei, Taiwan, ROC; Department of Orthopedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, ROC; Institute of Clinical Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC. Electronic address: drwuvhg@gmail.com.
⁸ Department of Orthopedics, Taipei Veterans General Hospital, Taipei, Taiwan, ROC; Department of Orthopedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, ROC. Electronic address: wmchen@vghtpe.gov.tw.
⁹ Institute of Traditional Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC; Department of Orthopedics, Taipei Veterans General Hospital, Taipei, Taiwan, ROC; Department of Orthopedics, Therapeutical and Research Center of Musculoskeletal Tumor, Taipei Veterans General Hospital, Taipei, Taiwan, ROC. Electronic address: yollywang@gmail.com.

PMID: 37321426
DOI: 10.1016/j.jep.2023.116784

Abstract

Ethnopharmacological relevance: The treatment of osteoarthritis (OA) patients is a challenging problem. Mesenchymal stem cells (MSCs) are multipotent cells and play key roles in regenerative medicine for cartilage degeneration. GuiLu-ErXian Glue (GLEXG) is an herbal remedy widely used in traditional Chinese medicine to treat joint pain and disability in elderly OA patients. However, the mechanisms of how GLEXG affects MSCs-induced chondrogensis remains to be elucidated.

Aim of the study: The aim of this study was to investigate the effects of GLEXG on MSC-derived chondrogenesis, both in vitro and in vivo and its potential mechanisms.

Methods: Using human MSC (hMSCs) as in vitro model, the effects of HPLC-profiled GLEXG water extract on chondrogenic differentiation were investigated by 3D spheroid cultures under chondrogenesis-inducing medium (CIM) condition. The chondrogenesis process was evaluated by measuring the sphere sizes, chondrogenesis-related genes expression by reverse transcription real-time PCR that targeted type II/X collagens, SOX9, aggrecan, and protein expression by immunostaining. Anti-TGF-β1 neutralization antibody was used for mechanistic study. Mono-iodoacetate (MIA) induced OA joint was used to evaluate the effects of GLEXG on in vivo model. MSCs-derived exosomes were purified for proteomics study and senescence process was evaluated by cumulative population doublings and senescence-associated β-Galactosidase staining.

Results: The results showed that GLEXG enhanced hMSCs chondrogenesis and upregulated RNA expression of type II/X collagen, SOX9 and aggrecan at 0.1 μg/mL, 0.3 μg/mL in vitro. In vivo, GLEXG at the dose of 0.3 μg intraarticular (i.a.) injection rescued the MIA-induced cartilage defect. Proteomics and ingenuity pathway analysis obtained from MSCs-released exosomes suggested that senescence pathway was less activated in GLEXG group than in vehicle group. Besides, GLEXG was able to increase cumulative population doubling and delayed hMSCs senescence process after four passages in cultures.

Conclusion: we conclude that GLEXG promotes in vitro MSC-induced chondrogenesis possibly via exosomes release and delays aging in the MSC senescence process and that treatment with GLEXG (0.3 μg, i.a.) rescued cartilage defects in rat OA knee model.

Keywords: Chondrogenesis; Exosome; GuiLu-ErXian glue; Herbal medicine; Mesenchymal stem cells; Osteoarthritis.

MeSH terms

Aged
Aggrecans / genetics
Aggrecans / metabolism
Aggrecans / pharmacology
Aging
Animals
Cell Differentiation
Cells, Cultured
Chondrogenesis / genetics
Collagen Type II / metabolism
Collagen Type X / metabolism
Exosomes* / metabolism
Humans
Mesenchymal Stem Cells*
Osteoarthritis*
Rats

Substances

Aggrecans
Collagen Type II
Collagen Type X